Task

#A verifiable and long read length NGS sequencing method (from chemistry, sequencer, to algorism)

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## Fig1 intro 2+2

* Work flow

* 1x4 work flow

* 2+2 work flow

* KEY:没有空cycle,homopolymers更长。

* Verifiable

* 010101XOR清晰的校验规则

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##Fig2 Fluorogenic sequencing demonstration

* Core fluorogenic chemistry (on surface) . KEY:突出化学

* TG-TPLFNS. KEY:突出化学

* TG spectra specs. KEY:突出化学

* Key kinetic

* homopolymer聚合酶的动力学方程

* data: single TPLFN and double TPLFNs mix. KEY:一种碱基两种碱基动力学并无区别;homopolymer时间上无区别

* Sequencer, a complete sequencing work flow (画个更好的版本)

* 仪器结构

* 每轮反应中的真实信号,温度等等

* Flow Chip, Surface and PCR. KEY: 突出化学工作

* how to achieve in high throughput format

* Surface stable test. Signal degeneration

* Sample data => sequence.  KEY:前面说的都是可行的:

* 1x4

* 2+2

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##Fig3 Long read length (from same time)

* Table:1x4 and 2+2 comparison(infinite length random sequence  simulation)

* Signal cycle rate

* Homopolymer  distribution

* Effective cycle before degeneration

* Carry Forward and IE

* 模拟CAFIE的结果(重要)

* 特征:长程累积,越展越开。一定限度就不提供信息量了。

* 一个过的去,两个过不去

* CF+IE+DG algorism: simulation, illustration of carry forward and insufficient extension.

* 中间出错不影响后面矫正。

* 可以翻一倍左右有效读长

* Corrected data

* Typical CF, IE degeneration value.

* Bar_nt: RAW => corrected , long readlength

* Table: Fluorogenic Sequencing 1x4, 2+2, RAW, after correction

* Table: 在不同的CFIE参数下,不同测序仪可达到的读长。KEY: 隐含可以普适。

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##Fig4 Verifiable (accuracy)

* Information calculation table (redundant information)

* Rectification

* False positive chart: Very low FP compare to 1x4

* Table: 在不同的错误率下,可达到的准确度。

* 不同homopolymer长度的indel 准确度。

* Substitution的比较。

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##Fig5 Best results

* Table:sample,HP分布,readlength,errorrate,FP,等等。

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